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从玉米(L.)幼苗衍生的分裂节外植体高效直接诱导芽器官发生。

Efficient direct shoot organogenesis from seedling derived split node explants of maize ( L.).

作者信息

Mushke Ramesh, Yarra Rajesh, Bulle Mallesham

机构信息

Centromere Bio solutions Pvt. Ltd, Vibha Seeds Group, Agri Science Park, ICRISAT, Patancheru, Hyderabad, Telangana State, India.

Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad, Telangana State 500046, India.

出版信息

J Genet Eng Biotechnol. 2016 Jun;14(1):49-53. doi: 10.1016/j.jgeb.2016.03.001. Epub 2016 Apr 8.

Abstract

Maize is one of the important cereal crops around the world. An efficient and reproducible regeneration protocol direct organogenesis has been established using split nodes as ex-plants derived from 7 to 10 day old grown seedlings. Surface sterilized maize seeds were germinated on MS medium lacking plant growth regulators. Nodal sections of 7-10 day old seedlings were isolated, split longitudinally into two halves and cultured on regeneration medium containing different concentrations of 6-benzyladenine (2.20, 4.40, 6.60, 8.80, 11.0 and 13.2 μM) or kinetin (2.32, 4.65, 6.97, 9.29, 11.6 and 13.9 μM). Inclusion of 8.80 μM BA into MS supplemented medium triggered a high frequency of regeneration response from split node explants with a maximum number of shoots (12.0 ± 1.15) and the highest shoot length (3.0 ± 0.73) was obtained directly (without an intervening callus phase) within 4 weeks of culture. Further shoot elongation was achieved on medium containing 4.40 μM BA. The elongated micro shoots were rooted on MS medium fortified with 1.97 μM indole-3-butyric acid. The regenerated plantlets with roots were successfully hardened on earthen pots after proper acclimatization under greenhouse conditions. This new efficient regeneration method provides a solid foundation for genetic manipulation of maize for biotic and abiotic stresses and to enhance the nutritional values.

摘要

玉米是世界上重要的谷类作物之一。利用7至10日龄幼苗的分裂节作为外植体,建立了一种高效且可重复的直接器官发生再生体系。将表面消毒的玉米种子在不含植物生长调节剂的MS培养基上萌发。分离7至10日龄幼苗的节段,纵向切成两半,接种在含有不同浓度6-苄基腺嘌呤(2.20、4.40、6.60、8.80、11.0和13.2 μM)或激动素(2.32、4.65、6.97、9.29、11.6和13.9 μM)的再生培养基上。在添加MS的培养基中加入8.80 μM BA可引发分裂节外植体的高频再生反应,在培养4周内直接获得最多的芽数(12.0±1.15)和最高的芽长(3.0±0.73)(无愈伤组织阶段)。在含有4.40 μM BA的培养基上进一步促进芽伸长。伸长的微芽在添加1.97 μM吲哚-3-丁酸的MS培养基上生根。生根的再生植株在温室条件下适当驯化后,成功地在花盆中炼苗。这种新的高效再生方法为玉米针对生物和非生物胁迫的遗传操作以及提高营养价值提供了坚实的基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/691b/6299849/144a429c1d65/gr1.jpg

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